AU2010324437A1 - Depolymerized glycosaminoglycan from Thelenota ananas and preparation method thereof - Google Patents
Depolymerized glycosaminoglycan from Thelenota ananas and preparation method thereof Download PDFInfo
- Publication number
- AU2010324437A1 AU2010324437A1 AU2010324437A AU2010324437A AU2010324437A1 AU 2010324437 A1 AU2010324437 A1 AU 2010324437A1 AU 2010324437 A AU2010324437 A AU 2010324437A AU 2010324437 A AU2010324437 A AU 2010324437A AU 2010324437 A1 AU2010324437 A1 AU 2010324437A1
- Authority
- AU
- Australia
- Prior art keywords
- dthg
- glycosaminoglycan
- depolymerized
- thg
- fgag
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 229920002683 Glycosaminoglycan Polymers 0.000 title claims abstract description 47
- 241000941620 Thelenota ananas Species 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title description 11
- 238000000034 method Methods 0.000 claims abstract description 52
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 claims abstract description 49
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 42
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 claims abstract description 28
- 150000002978 peroxides Chemical class 0.000 claims abstract description 20
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims abstract description 19
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 claims abstract description 19
- 229940097043 glucuronic acid Drugs 0.000 claims abstract description 19
- 208000007536 Thrombosis Diseases 0.000 claims abstract description 18
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 17
- 239000003054 catalyst Substances 0.000 claims abstract description 11
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 claims abstract description 6
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims abstract description 5
- 150000003467 sulfuric acid derivatives Chemical class 0.000 claims abstract 2
- 150000008267 fucoses Chemical class 0.000 claims description 21
- 150000003839 salts Chemical class 0.000 claims description 20
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 19
- 229940079593 drug Drugs 0.000 claims description 11
- 239000008194 pharmaceutical composition Substances 0.000 claims description 11
- 239000003814 drug Substances 0.000 claims description 10
- 229910021645 metal ion Inorganic materials 0.000 claims description 10
- 230000002265 prevention Effects 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000012736 aqueous medium Substances 0.000 claims description 4
- 239000011734 sodium Substances 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical group C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 159000000007 calcium salts Chemical class 0.000 claims description 2
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 230000000737 periodic effect Effects 0.000 claims 2
- 229910052723 transition metal Inorganic materials 0.000 claims 2
- 150000003624 transition metals Chemical class 0.000 claims 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 230000002785 anti-thrombosis Effects 0.000 abstract description 9
- 229910001428 transition metal ion Inorganic materials 0.000 abstract description 5
- 108010014173 Factor X Proteins 0.000 abstract description 4
- 239000012535 impurity Substances 0.000 abstract description 4
- 230000002429 anti-coagulating effect Effects 0.000 abstract description 3
- 230000003389 potentiating effect Effects 0.000 abstract description 2
- OVRNDRQMDRJTHS-KEWYIRBNSA-N N-acetyl-D-galactosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-KEWYIRBNSA-N 0.000 abstract 2
- 229940125532 enzyme inhibitor Drugs 0.000 abstract 1
- 239000002532 enzyme inhibitor Substances 0.000 abstract 1
- 229920000642 polymer Polymers 0.000 abstract 1
- 230000000694 effects Effects 0.000 description 62
- 238000006243 chemical reaction Methods 0.000 description 43
- 239000000047 product Substances 0.000 description 29
- 108090001015 cancer procoagulant Proteins 0.000 description 27
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 26
- 239000003146 anticoagulant agent Substances 0.000 description 25
- 239000000126 substance Substances 0.000 description 23
- 108090000481 Heparin Cofactor II Proteins 0.000 description 19
- 102000004032 Heparin Cofactor II Human genes 0.000 description 19
- 229940127219 anticoagulant drug Drugs 0.000 description 19
- 239000003055 low molecular weight heparin Substances 0.000 description 19
- 239000000243 solution Substances 0.000 description 19
- 238000001514 detection method Methods 0.000 description 18
- 230000001939 inductive effect Effects 0.000 description 18
- 229940127215 low-molecular weight heparin Drugs 0.000 description 18
- 230000001419 dependent effect Effects 0.000 description 17
- 239000000523 sample Substances 0.000 description 17
- 235000002639 sodium chloride Nutrition 0.000 description 17
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 17
- 241000251511 Holothuroidea Species 0.000 description 16
- 230000004071 biological effect Effects 0.000 description 16
- 238000005481 NMR spectroscopy Methods 0.000 description 14
- 239000004019 antithrombin Substances 0.000 description 13
- 230000014508 negative regulation of coagulation Effects 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 210000004369 blood Anatomy 0.000 description 12
- 239000008280 blood Substances 0.000 description 12
- 230000002401 inhibitory effect Effects 0.000 description 12
- 238000011160 research Methods 0.000 description 12
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 11
- 230000019635 sulfation Effects 0.000 description 11
- 238000005670 sulfation reaction Methods 0.000 description 11
- 241000335028 Lymantria grisea Species 0.000 description 10
- 230000005764 inhibitory process Effects 0.000 description 10
- 239000002244 precipitate Substances 0.000 description 10
- 230000009467 reduction Effects 0.000 description 10
- 238000004611 spectroscopical analysis Methods 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 208000032843 Hemorrhage Diseases 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 241000965254 Apostichopus japonicus Species 0.000 description 8
- 241001213462 Holothuria leucospilota Species 0.000 description 8
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- 230000000740 bleeding effect Effects 0.000 description 8
- 238000000605 extraction Methods 0.000 description 8
- 230000000144 pharmacologic effect Effects 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 241000258161 Stichopus Species 0.000 description 6
- 238000005100 correlation spectroscopy Methods 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 210000001835 viscera Anatomy 0.000 description 6
- 206010062713 Haemorrhagic diathesis Diseases 0.000 description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 description 5
- 206010047249 Venous thrombosis Diseases 0.000 description 5
- 239000003513 alkali Substances 0.000 description 5
- 239000013068 control sample Substances 0.000 description 5
- 150000002148 esters Chemical class 0.000 description 5
- 208000031169 hemorrhagic disease Diseases 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 235000011056 potassium acetate Nutrition 0.000 description 5
- 239000000376 reactant Substances 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 241000894007 species Species 0.000 description 5
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 4
- 244000184734 Pyrus japonica Species 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000004042 decolorization Methods 0.000 description 4
- 238000012691 depolymerization reaction Methods 0.000 description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 4
- 230000010118 platelet activation Effects 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 125000001424 substituent group Chemical group 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 3
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 3
- 229920001287 Chondroitin sulfate Polymers 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 241000258195 Holothuria Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 108090000190 Thrombin Proteins 0.000 description 3
- 102100030951 Tissue factor pathway inhibitor Human genes 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000003149 assay kit Methods 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 239000003729 cation exchange resin Substances 0.000 description 3
- 229940059329 chondroitin sulfate Drugs 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229920000669 heparin Polymers 0.000 description 3
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 3
- 108010013555 lipoprotein-associated coagulation inhibitor Proteins 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 230000003285 pharmacodynamic effect Effects 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 210000004623 platelet-rich plasma Anatomy 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000010254 subcutaneous injection Methods 0.000 description 3
- 239000007929 subcutaneous injection Substances 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- HRQDDZWMEGEOOO-UHFFFAOYSA-N 2-trimethylsilylpropanoic acid Chemical compound OC(=O)C(C)[Si](C)(C)C HRQDDZWMEGEOOO-UHFFFAOYSA-N 0.000 description 2
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000258955 Echinodermata Species 0.000 description 2
- 108010054218 Factor VIII Proteins 0.000 description 2
- 102000001690 Factor VIII Human genes 0.000 description 2
- 108010074860 Factor Xa Proteins 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 2
- 241000941624 Holothuria atra Species 0.000 description 2
- 238000004566 IR spectroscopy Methods 0.000 description 2
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 2
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 229910052783 alkali metal Inorganic materials 0.000 description 2
- 150000001340 alkali metals Chemical class 0.000 description 2
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000013522 chelant Substances 0.000 description 2
- 239000002738 chelating agent Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000009190 disseminated intravascular coagulation Diseases 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 229960000301 factor viii Drugs 0.000 description 2
- 230000020764 fibrinolysis Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 2
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229960003330 pentetic acid Drugs 0.000 description 2
- 229960001412 pentobarbital Drugs 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- -1 potassium acetate Chemical compound 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 229910021653 sulphate ion Inorganic materials 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 206010043554 thrombocytopenia Diseases 0.000 description 2
- 238000001551 total correlation spectroscopy Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 description 1
- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 description 1
- XNCSCQSQSGDGES-UHFFFAOYSA-N 2-[2-[bis(carboxymethyl)amino]propyl-(carboxymethyl)amino]acetic acid Chemical compound OC(=O)CN(CC(O)=O)C(C)CN(CC(O)=O)CC(O)=O XNCSCQSQSGDGES-UHFFFAOYSA-N 0.000 description 1
- NHQDETIJWKXCTC-UHFFFAOYSA-N 3-chloroperbenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 description 1
- 239000004342 Benzoyl peroxide Substances 0.000 description 1
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical compound C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229920000045 Dermatan sulfate Polymers 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- AFSDNFLWKVMVRB-UHFFFAOYSA-N Ellagic acid Chemical compound OC1=C(O)C(OC2=O)=C3C4=C2C=C(O)C(O)=C4OC(=O)C3=C1 AFSDNFLWKVMVRB-UHFFFAOYSA-N 0.000 description 1
- ATJXMQHAMYVHRX-CPCISQLKSA-N Ellagic acid Natural products OC1=C(O)[C@H]2OC(=O)c3cc(O)c(O)c4OC(=O)C(=C1)[C@H]2c34 ATJXMQHAMYVHRX-CPCISQLKSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010048049 Factor IXa Proteins 0.000 description 1
- 108010071289 Factor XIII Proteins 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- 206010062506 Heparin-induced thrombocytopenia Diseases 0.000 description 1
- 101000635804 Homo sapiens Tissue factor Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010023237 Jugular vein thrombosis Diseases 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 101100505672 Podospora anserina grisea gene Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000887536 Stichopus horrens Species 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- AVJBPWGFOQAPRH-FWMKGIEWSA-N alpha-L-IdopA-(1->3)-beta-D-GalpNAc4S Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS(O)(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C(O)=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 230000001858 anti-Xa Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000000702 anti-platelet effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 210000000702 aorta abdominal Anatomy 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000019400 benzoyl peroxide Nutrition 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 229960000182 blood factors Drugs 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- NEUSVAOJNUQRTM-UHFFFAOYSA-N cetylpyridinium Chemical compound CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 NEUSVAOJNUQRTM-UHFFFAOYSA-N 0.000 description 1
- 229960004830 cetylpyridinium Drugs 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- OPQARKPSCNTWTJ-UHFFFAOYSA-L copper(ii) acetate Chemical compound [Cu+2].CC([O-])=O.CC([O-])=O OPQARKPSCNTWTJ-UHFFFAOYSA-L 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- MRIZMKJLUDDMHF-UHFFFAOYSA-N cumene;hydrogen peroxide Chemical compound OO.CC(C)C1=CC=CC=C1 MRIZMKJLUDDMHF-UHFFFAOYSA-N 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940051593 dermatan sulfate Drugs 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229940012444 factor xiii Drugs 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 125000002446 fucosyl group Chemical group C1([C@@H](O)[C@H](O)[C@H](O)[C@@H](O1)C)* 0.000 description 1
- 125000002367 glucuronosyl group Chemical group 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 208000014951 hematologic disease Diseases 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 238000003929 heteronuclear multiple quantum coherence Methods 0.000 description 1
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 229940029329 intrinsic factor Drugs 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 210000004731 jugular vein Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 150000001455 metallic ions Chemical class 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- MGFYIUFZLHCRTH-UHFFFAOYSA-N nitrilotriacetic acid Chemical compound OC(=O)CN(CC(O)=O)CC(O)=O MGFYIUFZLHCRTH-UHFFFAOYSA-N 0.000 description 1
- 239000012299 nitrogen atmosphere Substances 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 230000033885 plasminogen activation Effects 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000001374 small-angle light scattering Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- AYRVGWHSXIMRAB-UHFFFAOYSA-M sodium acetate trihydrate Chemical compound O.O.O.[Na+].CC([O-])=O AYRVGWHSXIMRAB-UHFFFAOYSA-M 0.000 description 1
- 229940087562 sodium acetate trihydrate Drugs 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 229960003766 thrombin (human) Drugs 0.000 description 1
- 230000002537 thrombolytic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/737—Sulfated polysaccharides, e.g. chondroitin sulfate, dermatan sulfate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- General Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Disclosed is a depolymerized glycosaminoglycan from Thelenota ananas(dTHG), weight average molecular weight of which is about 8000 ~ 20000Da, and monosaccharide components of which are acetylgalactosamine(GalNAc), glucuronic acid(GlcUA), fucose(Fuc) or their sulfates(expressed as -OSO ), in which molar ratio of GalNAc: GlcUA: Fuc: -OSO is about 1: (1±0.3 ): (1±0.3): (3.5±0.5). Said dTHG is a potent endogenous factor X enzyme inhibitor, which has good anticoagulating and antithrombotic activity, and can be used to prevent and/or treat thrombotic diseases. Also provided is a method for preparing said dTHG, which comprises steps of 1) extracting and obtaining fucosylated glycosaminoglycan(THG) from Thelenota ananas body wall; 2) depolymerizing THG to obtain dTHG by method of peroxide depolymerization or method of peroxide depolymerization catalyzed by catalyst of the fourth period transition metal ions; 3) removing oligomer and/or polymer impurities in dTHG.
Description
Depolymerized Glycosaminoglycan from Thelenota ananas and Preparation Method Thereof FIELD OF THE INVENTION The present invention relates to the field of medical technology, and particularly to a depolymerized glycosaminoglycan from Thelenota ananas and a preparation method of the same, a pharmaceutical composition containing the same and use of the same in preventing and/or treating thrombotic diseases. The present invention is a further invention based on Chinese patent application No. 200910110114.0 entitled "Depolymerized Fucosylated Glycosaminoglycan and Preparation Method of the same" filed by the same applicant on November 6, 2009, the entire content of such application is incorporated herein by reference. BACKGROUND OF THE INVENTION Fucose-branched glycosaminoglycan (fucosylated glycosaminoglycan or fucose-containing glycos-aminoglycan, FGAG), also referred to as fucose-branched chondroitin sulfate (FCS), is a special glycosaminoglycan having sulfated fucose substituents extracted from the body wall or viscera of an echinoderm (Huizeng Fan, et al., Pharmaceutical Journal, 1980, 18(3): 203; Ricardo P. et al., J. Biol. Chem., 1988, 263 (34) : 18176; Yutaka K. et al., J. Biol. Chem., 1990, 265:5081). The exsiting literature shows that FGAG from sea cucumber have both common properties and differences. First, FGAG from different sources and prepared by different methods have some common features, i.e., FGAG has monosaccharide components including N-acetyl-galactosyl (GalNAc, A, chemical name of said N-acetyl-galactose is N-acetyl-2-deoxy-2-galactosamine, hereafter the same), glucuronosyl (GlcUA, U), fucosyl (Fuc, F) and their sulphate (see above reference of Huizeng Fan, 1980; Ricardo P, 1988; and Ken-ichiro Y. et al., Tetrahedron Letters, 1992, 33(34): 4959). In which, GlcUA and GalNAc (or its sulphate) interconnects through P (1-3) and P (1-4) glycosidic bonds to form a backbone, which is similar to a [GlcUA p(1-3)- GalNAc p(1-4)-] disaccharide repeating structural unit of chondroitin sulfate, while fucose or its sulfate attached to the backbone as a branch. FGAG from different sources and prepared by different methods have chemical structural differences to varying degrees, for example: 3395121_1 (GHMatters) P90582.AU (1) Difference in monosaccharide component ratio. FGAG from different sea cucumber species, from different tissues and even prepared by different methods may have significant difference in monosaccharide components. The monosaccharide components of FGAG from several sources are shown in Table 1. Table 1 Monosaccharide components and their sulfate groups of FGAG from several sea cucumber Sources Chemical components (molar ratio) References Sea cucumber species Tissues A: U: F: -OS0 3 Huizeng Fan, Pharmaceutical Journal, Stichopus aponicus 1:1.14 : 0.97 :4.20 1980,15: 267 1 :0.88: 0.93 :4.01 Ken-ichiro Y, Tetrahedron Letters, 1992, 33: body 4959 wall 1 :1.18 : 2.78: 5.46 Yutaka K, J. Biol. Chem., 1990, 265: 5081 1 :0.84:2.38 : 3
.
6 9 bI Yutaka K, Biochem. J, 2002,132: 335 viscera 1 :1.00:1.00 :4.70 Huizeng Fan, marine drugs, 1983, (3): 134 Stichopus variegatus body 1 1.21: 1.29: 4.62 Judi Chen, Chinese Journal ofMarine wall Drugs, 1994, (1). 24 Holothuria : 0.94 0.84 3.60 Huizeng Fan, Pharmaceutical Journal, leucospilota body 1983,18 (3): 203 wall : 0.96 0.78 1.98 Haitang Li, Journal of Chinese Medicinal Materials, 1999, 22(7):328 Holothuria atra body 1 :1.15 : 0.79 : 2.7 (el Xiaoli Tang, Journal of Chinese Medicinal wall Materials, 1999, 22(5):223 Holothuria scabr body 1 1.28: 0.68 : 1.72 Jian Chen, Food and Fermentation wall Industries, 2006, 32: 123 Ludwigothurea grisea 1 :1.17: 2.17 : 2.39 'c Paulo AS, Eur. J Biochem., 1987, 166:639 body 1: 0.90 : 0.97 :2.67 [d] Ricardo P V, J. Biol. Chem., 1988, 263: wall 18176 1 :0.92: 1.23 :2.21 Paulo AS, J. Biol. Chem., 1996, 271:23973 jai expressed as mmol/g in the reference (0.81:0.69:1.93:2.99); lb expressed as wt% in the reference (16.2: 20.3: 11.66: 23.52); Ic1 expressed as 0.46 : 0.54 : 1.00 : 1.10 in the reference; [dl expressed as 0.33 : 0.30 : 0.32 : 0.88 in the reference. The difference between the components of FGAG obtained from different species, tissue origins and different extraction methods mainly lies in the larger change in the ratio of Fuc to sulfate group. It can be judged based on the data that not only different specie sources result in different FGAG, but also different extraction methods may lead to a larger difference in components of the product. For example, Yutaka K et al. (1990, 2002), Huizeng Fan et al. (1980) and Ken-ichiro Y et.al (1992) respectively extracted FGAG from the body wall of 2 33951211 (GHMatters) P90562AU Stichopus Japonicus, however the molar ratio of fucose in FGAG obtained by the former was 2 to 3 times higher than that of the latters; while FGAG extracted from the same sea cucumber species (L.grisea) by the same research group also have larger differences in the ratio of fucose component (Paulo AS et al., 1987, 1988). FGAG obtained from the same tissue by different extraction method/time have different ratios of fucose component, it is probably because there exists non-GAG like fucosan pollution in FGAG product with higher ratio of fucose component, or there exists damage and loss of branch groups during the production of FGAG product with lower ratio of fucose component; in addition, it is probably related to the accuracy of content determination method. (2) Difference in structure of backbone: As there are differences in the position and number of sulfate groups of such as chondroitin sulfate A, C and D, the differences in the position and number of sulfate groups also exist in the backbone of FGAF from different species. For example, the data show that GalNAc on the backbone of FGAG from Stichopusjaponicas is sulfated at both 4- and 6-positions (Ken-ichiro Y et al., Tetrahedron Letters, 1992, 33: 4959); GalNAc on the backbone of FGAG from Holothuria leucospilota is sulfated only at 6-position but not 4-position (Huizeng Fan, Pharmaceutical Journal, 1983, 18 (3): 203); and on the backbone of FGAG from L. grisea, about 53% of the GalNAc are 6-sulfated, and small amounts of 4,6-sulfated (about 12%), 4-sulfated (about 4%) and nonsul fated (31%) are found (Lubor Borsig et al. J. Biol. Chem. 2007, 282: 14984). (3) Difference in fucose residues on branch and their sulfation degree. Data show that FGAG from both S. Japonicus and L. grisea have three types of fucose residue branches, i.e., 2,4-disulfated, 3,4-disulfated, and 4-monosulfated fucoses; and the latter has about 25% of nonsulfated fucose residues, which occur as a cluster at the reducing end of FGAG (Ken-ichiro Y. et al., Tetrahedron Letters, 1992, 33: 4959, Paulo AS. et al., J. Biol. Chem., 1996, 271: 23973). As shown in Table 1, FGAG from Stichopusjaponicus and Stichopus variegates generally show a higher degree of sulfation, while FGAG from L. grisea, Holothuria atra, and Holothuria scabr show lower degree of sulfation. The exsiting data shows that FGAG from echinoderm has various biological activities. Most FGAG from various sources has certain anticoagulant activity (Huizeng Fan, Pharmaceutical Journal, 1980, 15(5): 263;Peiwen Zhang, Chinese Journal of Pharmacology and Toxicology, 1988, 2(2): 98; Paulo AS. et al., J. Biol. Chem. 3 3395121_1 (GHMatters) P90582.AU 1996, 271: 23973); however, these natural FGAG also has platelet aggregation inducing acticity (Jia-zeng L. et al, Thromb Haemos, 1988, 54(3): 435; Chunwen Shan, Pharmacology and Clinics of Chinese Materia Medica, 1989, 5(3): 33). FGAG has also been reported to have biological activities of regulation of blood lipid (His-Hisen L. et al., J. Agric. Food Chem., 2002, 50: 3602), anti-artery atherosclerosis, and inhibition of vascular endothelial proliferation (Tapon Bretaudiere et al., Thromb. Haemost, 2000, 84: 332; Masahiko I. et al., Atherosclerosis, 1997, 129:27; European patent application, EP 0811635), immunoregulation (Ling Sun et al., Progress in biochemistry and biophysics, 1991, 18 (5): 394; Zuqiong Chen et al., Tianjin Medical Journal, 1987, (5): 278), antineoplastic (Renjie Hu et al., Chinese Journal of Clinical Oncology, 1992, 19(1): 72; Weimin Li et al., Journal of Clinical Oncology, 1985, 12(2): 118) and antivirus (JA. Beutler et al., Antivir. Chem. Chemother., 1993, 4(3), 167; PCT patent application PCT/JP90/00159), and so on. Research data about antithrombotic activity, anticoagulant activity and action mechanisms and pharmacological targets of FGAG from sea cucumber and its derivatives show that FGAG has an anticoagulant mechanism that is different from that of heparin and dermatan sulfate, the targets of its anticoagulant/antithrombotic effect may relate to: (1) AT-III: namely, there exists an AT-III-dependent antithrombin activity (Paulo AS et al., J. Biol. Chem., 1996, 271, 23973; Xi Ma, Chinese Journal of Hematology, 1990, 11(5):241); (2) HC-II: namely, there exists an HC-II-dependent antithrombin activity (Hideki Nagase et al., Blood, 1995, 85, (6):1527; Guangsen Zhang, Chinese Journal of Hematology, 1997, 18(3): 127); (3) IIa: namely, inhibiting feedback activation factor XIII of thrombin (Ila) (Nagase H et al., Biochem. J., 1996, 119(1): 63-69); (4) f.xase: inhibiting the activation of factor X by endogenous factor Xase (factor VIII-IX complex) (Hideki Nagase et al., Blood, 1995, 85(6):1527; JP Sheehan et al., Blood, 2006, 107(10): 3876); (5) TFPI: increasing the rate of factor Xa inhibition by TFPI, reducing the inhibitory activity on TF-factor VIla by TFPI-factor Xa, and stimulating the release of TFPI (Hideki Nagase et al., Thromb Haemost, 1997, 78: 864; T Bretaudiere et al., Thromb Haemost, 2000, 84: 332); (6) plasmin: promoting plasminogen activation and thus promoting thrombolysis (Xiaoguang Yang et al., Chinese Medical Sciences Journal, 1990, 12(3), 187; Yutaka Kariya et al., Biochem. J., 2002, 132. 335). 4 3395121_1 (GHMatters) P90502.AU Although FGAG from sea cucumber has an important potential application value due to its unique anticoagulant mechanism and good anticoagulant activity intensity, so far FGAG is difficult to be used in clinical and this is mainly because: (1) FGAG from sea cucumber has both anticoagulant activity and platelet aggregation-inducing activity. In clinical, the aim of anticoagulant is antithrombotic, and antiplatelet is another important approach to anti-thrombosis which is commonly used in clinical. Apparently, in terms of thrombosis, the anticoagulant activity for anti-thrombosis and the platelet-activating effect of promoting thrombosis of FGAG oppose each other, allowing it to be difficult to be used in clinical for preventing and treating hematologic diseases. For example, the studies indicate that in acute disseminated intravascular coagulation model of rabbits, the platelet-activating effect of FGAG from Stichopusjaponicas entirely offsets its anticoagulant efficacy (Anguo Li, Journal of Traditional Chinese Medicine University of Hunan, 1991, 11(3): 37). (2) Administration of pharmacodynamic dose of FGAG from Stichopus japonicas into the blood vessels of living animals can lead to platelet count reduction (Jiazeng Li, Chinese Pharmacological Bulletin, 1985, 6(2): 107). Thrombocytopenia, such as immune heparin-induced thrombocytopenia, may lead to hemorrhagic tendency or may lead to serious or even fatal disseminated intravascular coagulation. Available data show that platelet count reduction induced by FGAG from sea cucumber may be related to its platelet aggregation-inducing activity and thus "withhold" of the platelets in the microvascular (Jiazeng Li et al., Bulletin of Chinese Materia Medic, 1983, 8(5): 35). (3) It is generally known that a wide range of pharmacological targets are closely related to the side effect of bleeding tendency of anticoagulant, and selective target has become an important evaluation index in the development of new anticoagulant drugs (KA Bauer, Hematology, 2006, (1): 450). FGAG from sea cucumber has different targets from that of the other anticoagulant drugs, however, as mentioned above, its targets are still relatively widespread. Available data show that an anticoagulant dose of FGAG can lead to significant bleeding tendency (Paulo AS et al., British Journal of Haematology, 1998, 101: 647). Natural FGAG from sea cucumber has unique anticoagulant mechanism and potency, on the other hand has defects that limit its application. So, the acquisition of desired target products through structural modification has become one of the important contents of the application research. 5 3395121_1 (GHMatters) P90582.AU At present, the methods for the chemical structural modification of FGAG comprise peroxide depolymerization (European patent disclosure, EP0408770; Ken- ichiro Yet al., Tetrahedron Letters, 1992, 33: 4959), desulfation or carboxyl reduction (Paulo AS et al., Thrombosis Research, 2001, 102: 167), partial acid hydrolysis (Yutaka Kariya, Biochem. J., 2002, 132: 335; Paulo AS et al., Thrombosis Research, 2001, 102: 167), and etc.. These efforts have made some progress. For example, studies show that the platelet aggregation-inducing activity of FGAG from Stichopusjaponicas may weaken with the reduction of molecular weight (Huizeng Fan, Journal of Biological Chemistry, 1993, 9(2):146); AT-III-dependent antithrombin activity of the depolymerized product by peroxide of FGAG from Stichopusjaponicas may also be reduced (Xi Ma, Chinese Journal of Hematology, 1990, 11(5): 241 ; Paulo AS et al., J Biol. Chem. 1996, 271, 239 73; Hideki Nagase et al., Blood, 1995, 85 (6):1527); For FGAG from L. grisea, the reduction of molecular weight has a more significant effect on HC-II-dependent antithrombin activity (RG Pacheco et al., Blood Coagulation and Fibrinolysis, 2000, 11.563). By summarizing the research data about depolymerized product of FGAG (mostly referred to as DHG) from Stichopusjaponicas, it is known that it is difficult to obtain anticoagulant active products with desired potency and target feature. For example, the data show that the platelet aggregation-inducing activity may be eliminated until the weight-average molecular weight of FGAG from Stichopusjaponicus is reduced to 9000Da (Huizeng Fan et al., Journal of Biological Chemistry, 1993, 9(2): 146); on the other hand, the anticoagulant activity weakens with the reduction of molecular weight (RG Pacheco et al., Blood Coagulation and Fibrinolysis, 2000, 11:563; Huizeng Fan et al., Journal of Biological Chemistry, 1993, 9(2): 146). According to the DHG-related pharmacological and pharmacodynamic research data, DHG with molecular weight less than I OOOODa was used in early, but in the subsequent more than ten years of research, weight-average molecular weight of DHG is mostly between 12000 and 1 5000Da (Hideki Nagase et al., Thromb Haemost, 1997, 77(2): 399; Kazuhisa Met al., Kidney International, 2003, 63: 1548; JP Sheehan et al., Blood, 2006, 107(10): 3876). Apparently, the latter is required to maintain necessary anticoagulant potency, but for FGAG from Stichopus japonicas, the safety of this molecular weight range is in doubt in terms of eliminating platelet-inducing activity and 6 3395121_1 (GHMetters) P90502.AU avoiding intravascular administration-induced thrombocytopenia, and such safety has not been reflected and validated in the related research data. Studies have shown that both hydrolysis and partial desulfation of fucose branches can significantly reduce or abolish anticoagulant activity of FGAG, reduction of carboxyl groups has relatively less effect on anticoagulant activity, but the bleeding tendency is still apparent and the effect on platelet activity is unknown (Paulo AS, J. Biol. Chem., 1996, 271, 23973, Paulo AS et al., British J. Haematology, 1998, 101: 647, Paulo AS et al., Thrombosis Research, 2001, 102: 167). Research data on chemical and biological activity of FGAG from sea cucumber mainly relates to FGAG from L. grisea and Stichopus japonicas. The extraction method of glycosaminoglycan from Thelenota ananas has been described (Xuexiang Liuet al., Journal of Nanjing University of Traditional Chinese Medicine, 2003, 19 (3): 161), but studies on its structure analysis and biological activity have not been reported. It is seen from available data that the difference between structures of FGAG from L. grisea and Stichopusjaponicas lies in the degree of sulfation. GalNAc backbones have different sulfated types and levels, and fucose branches have substantially the same type; while different types of fucose branches have different compositions and thus have different degrees of sulfation on branches (Ken-ichiro Y et al., Tetrahedron Letters, 1992, 33: 4959; Lubor Borsig et al. J. Biol. Chem. 2007, 282: 14984). According to the relative anticoagulant potency of such two FGAG compared to heparin and/or low molecular weight heparin, it is known that FGAG from Stichopusjaponicas has stronger anticoagulant and antithrombotic potency (Norihiko S et al. Thromb Haemost, 1991, 65(4). 369; Paulo AS et al., British J Haematology, 1998, 101: 647). However, all the above efforts failed to illuminate the effect of platelet activation and intravenous administration of the obtained product on platelet count, which is the most important factor that limits the application value of FGAG from sea cucumber. Next, although the related documents reported the effect potency of these structure-modified products on certain blood factors (pharmacological targets), but the relationship between structural modification and features of pharmacological action mechanism of the obtained product is not clear. So far, the effect of sulfated position on the activity and action features of FGAG has not been reported. Alteration of sulfated type of the glycosyl groups on backbones and/or 7 3395121_1 (GHMalterm) P90582 AU branches may obtain a new FGAG having more selective pharmacodynamic characteristics and thus having more application values, however, under the existing conditions, glycosaminoglycan can be treated by nonselective sulfation or defulfation, but it is difficult to be modified by position-selective sulfation or defulfation. The present inventors find surprisingly by the comparative studies of the chemical structure and biological activity of FGAG from Thelenota ananas that the positions and types of fucose branches of FGAG have an important effect on the biological activity, especially on the platelet aggregation-inducing activity. Thus, FGAG from different species may have remarkable differences in application value. The present inventors also find that different depolymerization degrees and methods have different effects on the strength of biological effect that is produced by FGAG through different targets. Based on such difference, one can obtain the features with special target selectivity, and thus obtain FGAG derivatives for treating and/or preventing specific diseases. The present inventors first find that fucosylated glycosaminoglycan from Thelenota ananas (THG) has special chemical structure features, and its fucose branch types are different to that of FGAG with known or partially known chemical structure, such as FGAG from L. grisea and Stichopusjaponicas; THG also has special biological activities, and its platelet activation action is much lower than that of FGAG from such as Holothuria leucospilota and Stichopusjaponicas. The present invention demonstrates that THG has an activity of inhibiting endogenous factor X enzyme (f.Xase) and HC-II-dependent antithrombin(f.I]a), and first illuminates the relationship between the depolymerization degree of THG and the potency of the pharmacological action. Based on this, the present invention obtains depolymerized THG (dTHG) with higher ratio of f.Xase inhibition /anti Ia activity (potency ratio). Namely, based on the correlation rule between depolymerization and biological activity, the present invention obtains dTHG product with good anticoagulant potency and special target selectivity, starting from THG with special chemical and biological activity. Said dTHG has no platelet aggregation-inducing activity and does not cause platelet count reduction under conditions of high dose and repeated administration. SUMMARY OF THE INVENTION 8 3395121_1 (GHMatters) P90592 AU One object of the present invention is to provide a depolymerized glycosaminoglycan from Thelenota ananas (THG) with special chemical structure and good anticoagulant potency, which has no platelet aggregation-inducing activity and does not cause platelet count reduction. Another object of the present invention is to provide a method for preparing the depolymerized glycosaminoglycan from Thelenota ananas. Another object of the present invention is to provide a pharmaceutical composition containing the depolymerized glycosaminoglycan from Thelenota ananas, and its use for preparing medicines for prevention and/or treatment of thrombotic diseases. The present invention finds that besides some known common features of FGAG, such as monosaccharide components including GalNAc, GlcUA, Fuc and etc., glycosaminoglycan from Thelenota ananas (THG) has special chemical structure features. Its fucose branch type and sulfation degree are different from that of the known or partially known FGAG from sea cucumber. For example, FGAG from L. grisea and Stichopusjaponicas mainly comprise three types of fucose branches, i.e., -2,4-disulfated, -3,4-disulfated, and -4-sulfated fucoses, while THG mainly comprises -2,4-disulfated, -3-disulfated, and -4-sulfated fucoses. Accordingly, the present invention demonstrates that THG has the activity of inhibiting endogenous factor Xase (f.Xase) and HC-II-dependent antithrombin (f.IIa) and finds that THG exhibits special feature in biological activity, such as having a much lower activity in platelet activation than FGAG from Holothuria leucospilota and Stichopus japonicas. In addition, the present inventors find surprisingly by the comparative studies of the chemical structure and biological activity of THG that the positions and types of fucose branches of FGAG have an important effect on the biological activity, especially on the platelet aggregation-inducing activity. Based on the above researches, the present invention obtains depolymerized THG (dTHG) product with good anticoagulant potency and special target selectivity, starting from THG with special chemical and biological activity, based on the correlation rule between depolymerization and biological activity. According to one aspect of the present invention, said depolymerized THG (dTHG) is a depolymerized product of fucosylated glycosaminoglycan from Thelenota ananas (THG), having a structure shown in the following formula: 9 3395121.1 (GHMatters) P90582.AU O_ RO CH2 Ro RO 0 O- Oil Nil O (I) In formula (I): -OR represents hydroxyl groups (-OH), sulfate groups (-OSO 3 ), or sulfated fucose residues shown as formula (II) RO 0 RO 0 OR (II) In formula (II): -OR represents the same as formula (I). Wherein: monosaccharide components comprise N-acetyl galactosamine (GalNAc), glucuronic acid (GlcUA), fucose(Fuc), or sulfate thereof (-OSO3-), based on molar ratio, GalNAc: GlcUA: Fuc: -OS0 3 being about 1:(10.3):(I±0.3):(3.5±0.5); weight average molecular weight (Mw) of dTHG being about 8000-20000 Da. The molecular weight range is preferably 10000- 1 8000Da, and more preferably 12000-16000Da. Studies demonstrate that dTHG of the present invention has higher ratio of activity intensity (potency ratio) of anti f.Xase/anti f.IIa, higher potency ratio of antiXase/ APTT extension, has no platelet aggregation-inducing activity and does not cause platelet count reduction under conditions of high dose and repeated administration, and thus can be used for the prevention and/or treatment of thrombotic diseases. According to another aspect of the present invention, a method for preparing said depolymerized THG is provided, mainly comprising the following steps: 1) extraction: extracting and obtaining fucosylated glycosaminoglycan (THG) from the body wall of Thelenota ananas; 2) depolymerization: depolymerizing the THG obtained in step 1) to obtain depolymerized fucosylated glycosaminoglycan (dTHG); 3) purification: collecting and purifying dTHG with desired molecular weight to remove impurities with lower and/or higher molecular weight. 10 3395121_1 (GHMattesI) P90582 AU Particularly, step 1) generally may comprise the steps of cutting/grinding, enzymatic hydrolysis, alkaline hydrolysis, decolorization, separation. Thelenota ananas may be fresh or dry product without viscera. In order to increase the yield, said Thelenota ananas dried product is generally cut into flakes or small blocks and then immersed into water, while the fresh product may be cut and grinded directly by adding water into suspension, and then subjected to enzymatic and alkaline hydrolysis treatment. In the step of enzymatic hydrolysis, broad-spectrum protease is generally selected, including such as pepsin, trypsin, complex enzyme or crude enzyme from animal; prolease such as papain, actinase from plant and/or microorganism. The enzymatic hydrolysis conditions such as temperature, time, pH and amount of enzyme may be determined according to the properties of the used prolease. In the step of alkaline hydrolysis, strong alkali such as potassium hydroxide, sodium hydroxide may be used. Amount of alkali is selected to make the concentration of alkali in the extraction solution to be 0.5-2N (equivalent concentration), and reaction temperature may generally range from room temperature to 70'C, and alkali treatment time may be about 0.5-3h. The extraction solution after enzymolysis and/or alkali treatment is subjected to enzyme inactivation and neutralization, and then centrifugated and/or filtered to remove undissolved substances. The obtained supernatant may be treated or conjointly treated by a lower alcohol/ketone, such as ethanol, acetone; or a sylvite such as potassium acetate, or an acid mucopolysaccharide precipitant, such as cetylpyridinium precipitant to precipitate glycosaminoglycan in the solution. The obtained precipitant may be dried to get crude extract, or directly used in the next step without being dried. The obtained glycosaminoglycan crude extract may be purified by decolorization, fractional precipitation, gel permeation chromatography and/or ion exchange chromatography. Hydrogen peroxide decolorization combined with ehonal-potassium acetate fractional precipitation is preferably used in the present invention (see Huizeng Fan et al., Pharmaceutical Journal, 1983, 18 (3): 203) to obtain relatively purified fucosylated glycosaminoglycan from Thelenota ananas, i.e.THG. In prevent invention, fucosylated glycosaminoglycan from Thelenota ananas (THG) shares some common features with known fucosylated glycosaminoglycan, such as monosaccharide components including acetyl galactosamine (GalNAc), glucuronic acid (GlcUA ), fucose (Fuc) and etc., however, its degree of sulfation 33951211 (GHMatters) P90582 AU (expressed as -OSO ) is different from that of the known fucosylated glycosaminoglycan. Based on the integer (or half integer) molar ratio, the ratio of monosaccharide components and sulfate groups of GalNAc: GlcUA: Fuc: -OSO is close to 1: 1: 1: 3.5, and particularly about 1: (1±0.3): (1±0.3): (3.5±0.5). In step 2), depolymerization method of THG may select hydrogen peroxide depolymerization, or peroxide depolymerization catalyzed by the fourth period transition metal ions. The latter method is preferred in the present invention, which is described in detail in Chinese patent application No. 200910110114.0 entitled "Depolymerized Fucosylated Glycosaminoglycan And Preparation Method Thereof " submitted by the present applicant on November 6, 2009. It empolyes a catalyst containing the fourth period transition metal ions in aqueous medium to catalyze peroxide depolymerization and obtain a depolymerized fucosylated glycosaminoglycan from Thelenota ananas. Particularly, the method comprises the following steps: 2.1) adding peroxide in aqueous medium in the presence of the fourth period transition metal ions to depolymerize fucosylated glycosaminoglycan from Thelenota ananas (THG); 2.2) stopping the reation and collecting the depolymerized fucosylated glycosaminoglycan from Thelenota ananas (dTHG) within desired molecular weight range. Wherein, the depolymerization is carried out in aqueous medium, peroxide can produce free radicals in the reaction system and degrade THG through a free radical chain reaction to generate dTHG product. Said peroxide includes but not limited to peracetic acid, hydrogen peroxide, 3-chloro-peroxybenzoic acid, hydrogen peroxide cumene, sodium persulfate, benzoyl peroxide, and a salt or an ester thereof. Hydrogen peroxide is preferred. Said THG is about 0.05% to 15% based on the weight of the reaction system, and the peroxide is about 0.5% to about 30% based on the weight of the reaction system. During the depolymerization of FGAG, peroxide reactant may be added into the reaction system one-time before the reaction, and also may be added into the reaction system continuously or intermittently. Preferably, peroxide reactant is added into the reaction system continuously at a controlled rate in the present invention. Said metal ion as catalyst is selected from the fourth period transition metal ions, including such as Cu', Cu 2+, Fe 2+, Fe 3 *, Cr 3 *, Cr 2 07 2- Mn2+, Zn 2+, Ni 2 +. These 12 3395121_1 (GHMsfl8eS) P90582.AU metal ions may be used alone or combined with each other as composite catalyst, wherein preferred catalyst may include Cu', Cu 2+, Fe 2+, Fe 3, Zn 2+, and the most preferred is Cu 2 + Because metal ion is not a self-existent chemical reagent, actually used is an inorganic or organic salt of the metal ions. In the reaction system, the concentration of said metal may be about I nmol/L-O. 1 mol/L, and preferred concentration range is 10ptmol/L-10mmol/L. Said depolymerization reaction comprises the following conventional process parameters: temperature range is 10 C-75*C; reaction time is 20min-8h; the reaction may be carried out under atmospheric or pressurized conditions; may be carried out under inert gas such as nitrogen atmosphere, and also may be carried out under atmospheric conditions connected to atmospheric environment. When the reaction finishes, a chelating agent may be optionally added into the reaction system to chelate with the metal ions to inhibit reaction rate, then by cooling, organic solvent precipitant and other techniques to stop the reaction. Chelating agent is a substance that combines with a metallic ion to form an inert chelate, including but not limited to, ethylenediamine tetraacetic acid (EDTA), diethylenetriamine pentaacetic acid (DTPA), 3-propylenediamine tetraacetic acid (PDTA), nitrilotriacetic acid (NTA), or a salt thereof. Disodium ethylenediamine tetraacetic acid or its hydrate is preferred in the method of the present invention. Reaction product precipitation is a method that precipitates polysaccharides from the reaction system by directly adding an organic solvent or further adding an inorganic salt (such as potassium acetate). Said organic solvent includes low-carbon alcohol/ketone, such as methanol, ethanol and acetone, and preferably is ethanol or acetone. The above method can significantly improve the reaction condition of peroxide depolymerization of fucosylated glycosaminoglycan. That is, when employing the same peroxide reactant and original FGAG starting material to produce dFG with the same or approximately the same molecular weight and under the same temperature and other reaction conditions, this method can improve reaction rate, shorten reaction period, compared with direct peroxide depolymerization (namely the peroxide depolymerization method in the absence of metal ion catalyst). Similarly, under the condition of controlled reaction period, this method can significantly reduce the required reaction temperature, and even allow the reaction to proceed under room temperature. 13 3395121_1 (GHMotters) P90562 AU During the repeated preparation of dTHG, under the same or similar reaction conditions, compared with direct peroxide depolymerization, the differences between different batches of the products obtained by this method is significantly reduced, the difference between the molecular weight of obtained depolymerized product and target molecular weight is less than 5%, significantly improving the repeatability and controllability of dTHG production, and make the depolymerized product have better quality uniformity. This may be related to relatively stable reaction rate due to the decrease of activiation energy caused by catalyst, and may also be related to the moderate reaction conditions, such as lower reaction temperature and short reaction period, and the improvement of controllability and stability of reaction conditions. In addition, in order to further enhance and improve the controllability of the depolymerization reaction, an inorganic and/or organic salt at a certain concentration may optionally be added to the peroxide depolymerization reaction system catalysted by metal ions. Said inorganic and/or organic salts includes a salt formed by a metal element (such as an alkali metal, an alkaline earth metal element) and halogen or organic acid; a salt formed by an organic acid or inorganic acid with an organic base, and combinations thereof. Sodium chloride, potassium chloride, sodium acetate, trihydrate sodium acetate, potassium acetate are preferred. In the present invention, preferred concentration of the inorganic and/or organic salts used to improve depolymerization rate and reaction controllability is about 0.1 mmol/L to about 1.Omol/L. In the present invention, the collected dTHG has a weight average molecular weight (Mw) of about 8000-20000Da, preferably 10000- 1 8000Da, and more preferably 12000-16000Da. In step 3), the collected dTHG may be purified by a method known in the art to remove impurities with low and/or high molecular weight. Purification methods include but not limited to: dialysis method to remove low and/or high molecular weight impurities, ion-exchange method to produce a THG salt, and/or gel chromatography/anion exchange chromatography method. The method of preparing depolymerized fucosylated glycosaminoglycan from Thelenota ananas of the present invention may further comprise the following step: 14 3395121_1 (GHMailers) P90562.AU 4) drying the dTHG obtained in step 3). Drying methods may be decompression vacuum drying or freeze drying. Freeze drying is preferably used in the present invention. The ratio of monosaccharide components and sulfate groups of dTHG obtained by the above method of the present invention may be determined by the known methods in the art, such as chemical chromogenic method, infrared spectroscopy (IR) method, and nuclear magnetic resonance spectroscopy (NMR) method (Weijie Zhang, Biochemical Study Techniques of Glycoconjugates (Version 2), Zhejiang: Zhejiang University Press, 1999; Patent application of the present applicant mentioned above: CN200910110114.0). Since the dTHG of the present invention has sulfate groups and carboxyl groups, it can be used with an inorganic ion or an organic alkaline group to form a pharmaceutically acceptable salt or ester. Said pharmaceutically acceptable salt or ester of dTHG may be an alkali metal and/or alkaline earth metal salt, preferably sodium salt, potassium salt or calcium salt. Apparently, these salts or esters should also included within the scope of the present invention. According another aspect of the present invention, a pharmaceutical composition is provided, comprising dTHG of the present invention or a pharmaceutically acceptable salt or ester thereof, and pharmaceutically acceptable excipients. Said pharmaceutical composition may be formulated into various dosage forms, such as oral preparation (including solid and liquid preparation), injection preparation (including injection liquid and lyophilized powder for injection). However, the present inventors finds that of dTHG has poor oral absorption, but has good bioavailability when administrated by injection routes, such as subcutaneous injection. Thus, the pharmaceutical composition of the present invention is preferably formulated into injection preparation, and it can be formulated into a solution preparation and a freeze-dried product by conventional techniques in the art because dTHG has good water solubility. Said dTHG of the present invention is potent endogenous factor Xase inhibitor, and has good anticoagulant and antithrombotic activity, thus the above pharmaceutical composition containing dTHG can be used for the prevention and/or treatment of thrombotic diseases, such as the treatment of various postoperative anticoagulation, prevention and treatment of various arteriovenous thrombus, prevention and treatment of thrombus-related cardiovascular and I5 3395121_1 (GHMatters) P905e2.AU cerebrovascular diseases. Usage and dosage for the prevention and/or treatment of different diseases should be decided by clinical doctors. DESCRIPTION OF DRAWINGS Fig. 1: shows 'H NMR spectroscopies of THG and dTHG samples, wherein the water peaks in the spectroscopies of dTHG-6 and dTHG-9 are suppressed; Fig. 2: shows 'H- 1 H COSY spectroscopies of dTHG (A) and depolymerized FGAG from Stichopus Japonicus (i.e., dSJG, B); wherein signals indicated by arrows show the differences in the structure of fucose branches between the two depolymerized FGAG: Fig. A shows the related signals of 3-sulfated fucose of dTHG; Fig. B shows the related signals of 3,4-disulfated fucose of dSJG; signals in panes are respectively the related hydrogen signals of 3-sulfated fucose and 3,4-disulfated fucose; signals in the two circles are the signals of H-4 position of 3,4-disulfated fucose; Fig. 3: shows 3 C NMR and DEPT spectroscopies of THG and FGAG from Stichopus Japonicus (i.e., SJG); Fig. 4: NMR homonuclear/heteronuclear correlation spectroscopies of dTHG-6; Fig. 5: shows the relationship of the potency of the inhibition of f.Xase activity, HC-II-dependent anti-IIa acticity and the extension of APTT time, wherein the part defined in pane is the molecular weight of dTHG of the present invention; Fig. 6: shows the effect of dTHG and low molecular weight heparin (LMWH) on bleeding time in animals. DETAILED DESCRIPTION OF THE INVENTION The present invention can be further understood through the following detailed description of the examples in conjunction with the accompanying drawings. The examples should not be construed to limit the scope of the present invention. Example 1: Extaction, depolymerization and purification of THG 1.1 Materials Thelenota ananas, commercially available, viscera removal and body wall dried Stichopusjaponicas, commercially available, viscera removal and body wall dried H. leucospilota, commercially available, viscera removal and body wall dried. 16 3395121_1 (GHfttlters) P90582 AU Papain: 8x 10 5 U/g, available from Guangxi Nanning Pangbo Biological Engineering Co., Ltd. Cation exchange resin: 001 x7 strongly acidic styrene type cation exchange resin, available from Nankai University Resin Co. Ltd. (Tianjin). Reagents such as KOH, KCOCH 3 , H 2 0 2 , and ethanol were commercially available analytical reagents. 1.2 Extraction 20 kg dry Thelenota ananas were cut with slicer into sheets with a thickness of about 1.5 mm, placed into a laminated reaction vessel (300L), added with 200L water, stirred and soaked, and then added with solid NaOH under stirring until the concentration of NaOH reached 0.5M. Alkaline hydrolysis reaction was carried out at 604C for 2h. The reactants were cooled, adjusted to pH 6-7 with 6N HCl, and added with 1OOg of papain. The reaction was carried out at 50 C for 6h under stirring and then heated to 1 00 0 C maintaining for 1 0min. The reactants were cooled, adjusted to pH 2 with 6N HCl (to precipitate proteins), placed at 2-8'C for about 4h, and centrifugated to remove precipitate. The resultant supernatant was adjusted to about pH 7, added with ethanol until reaching a final concentration of 70%, and then still placed and centrifugated. The centrifuged precipitate was dissolved in 30 times (v/v) of water, centrifugated to remove undissolved substances, adjusted to about pH 10 with 2M NaOH, and added with H 2 0 2 until reaching a final concentration of about 3% (v/v), and then reacted at 50'C for 2h (decolourization). The reaction solution was added with potassium acetate until reaching a final concentration of 0.5mol/L, and added with ethanol until reaching a final concentration of 30%. The solution was then still placed and centrifugated. The centrifuged precipitate was dissolved in 20 times (v/w) of water, centrifugated to remove undissolved substances, added with potassium acetate until reaching a final concentration of 2.5mol/L, placed still and centrifugated. The centrifuged precipitate was washed twice with ethanol, and the residual ethanol was removed under reduced pressure. The precipitate was dissolved in water and then frozen-dried to obtain 162g of fucosylated glycosaminoglycan from Thelenota ananas (THG). As controls, 2kg of Stichopus Japonicus and Holothuria leucospilota were respectively extracted under the above conditions and 20.2g of FGAG from Stichopus Japonicus (SJG) and 15.6g of FGAG from Holothuria leucospilota (HLG) were obtained respectively. 17 3395121.1 (GHMatters) P90562.AU 1.3 Depolymerization and Purification 50g of THG obtained by the above method was added into 1825ml of water containing 122g of sodium acetate trihydrate and 60g of NaCl, and added with 120ml of 0.0668mol/L copper acetate solution, and mixed well under stirring. Under stirring condition in 35'C water bath, 10% (V/V) H 2 0 2 was dropwised at the rate of 126ml/h in about 2h. During the whole process, pH of the reaction system was controlled at 7.2-7.8. The reaction was carried out under above conditions for about 5h. During the reaction, about 180ml of reaction solution was taken at the point of 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5 and 5.Oh after the start of the reaction, respectively. Each of the reaction solution was added with 0.5g of Na 2 EDTA immediately after taking, adjusted to pH 6.5-7.0 with 15% acetic acid, then added with ethanol, 2.5 times the volume of the reaction solution (about 450ml), placed still and centrifugated to obtain precipitate. The precipitate was dissolved with 100ml of water, and precipitated again with ethanol (250ml of 95% ethanol). The centrifuged precipitate was washed twice with 50ml of ethanol, and ethonal was removed under reduced pressure. The precipitate was dissolved with 30 times volume (v/v) of water, filtrated through a 0.45pm membrane. The filtrate was passed through Na+ type cation exchange resin column (0 40 mm x 250 mm), the eluent was collected and dialyzed for 6h using a 3500-Da molecular weight cut-off dialysis membrane. The dialyzed cut-off solution was frozen-dried to obtain depolymerized products corresponding to each depolymeriztion time point dTHG-1~'dTHG-10. Amounts of product at each time point were about 3.0-3.5g, with a total amount of 33.2g. As controls, I Og SJG and 1 Og HLG were respectively taken and depolymerized under the same conditions as mentioned above. Two time points of taking the depolymerization reaction solution were set, i.e., 3h and 5h after the start of reaction. The obtained depolymerized products were designated as dSJG- 1, dSJG-2 and dHLG-1, dHLG-2. The total amount of dSJG was 7.5g, and the total amount of dHLG was 8.0g, 1.3 Determination and comparison of physical and chemical properties of dTHG, dSJG and dHLG Sample: THG, dTHG-lI--dTHG-10 Control sample: SJG, dSJG-1~dSJG-3; HLG; dHLG-1-dHLG-4 Detection of ,molecular weight: HPGPC-LALLS 18 33951211 (GHMaiers) P90582.AU Detection of optical rotation: Chinese pharmacopoeia (2005) part II, appendix VI E method. WZZ-1 S automatic polarimeter, sodium lamp (X 589.3nm), sample cell 1dm Detection of intrinsic viscosity: Chinese pharmacopoeia (2005) part II, appendix VI G, method 3 Detection of monosaccharide components: Acetylgalactosamine (GalNAc): Elson-Morgon method (Weijie Zhang, Biochemical Study Techniques of Glycoconjugates (Version 2), Zhejiang: Zhejiang University Press, 1999, 409-410) Glucuronic acid (GluUA): Carbazole method; Fucose (Fuc): cauculation of the molar ratio of GalNAc/Fuc according to integral area of methyl peaks of said 'H NMR in example 2 Sulfate group (-OSO3~): detection of the molar ratio of carboxylic acid/sulfuric acid by conductance method (Weijie Zhang, Biochemical Study Techniques of Glycoconjugates (Version 2), Zhejiang: Zhejiang University Press, 1999, 409-410) as the ratio of components GlcUA/-0S0 3 ~ Results: see Table 2. Table 2. Chemical components and physical and chemical properties of each sample Molecular Optical rotation Intrinsic viscosity Monosaccharide components Sample weight Opia2oain0nrni icst (molar ratio) (Mw) t , C=]%) (0.1M NaCl, ml/g) GaINAc: GcUA Fuc: -OSO3 THG 65820 -59.10 46.7 1.00 : 1.16 :1.01 : 3.60 dTHG-1 46070 -58.3* 29.3 1.00: 1.09 :1.00 : 3.52 dTHG-2 33260 -58.20 19.2 1.00 :1.12 :1.02 :3.61 dTHG-3 25380 -59.50 13.5 1.00 :1.03 : 0.98 : 3.45 dTHG-4 19650 -57.30 9.64 1.00: 1.06 :1.01 :3.50 dTHG-5 17150 -55.7* 8.07 1.00: 1.15 :0.97 :3.42 dTHG-6 13950 -53.7* 6.17 1.00 :1.03 : 0.96 : 3.53 dTHG-7 11580 -58.4* 4.84 1.00 :1.08 :1.00: 3.48 dTHG-8 10260 -55.90 4.12 1.00: 1.04 :0.99 :3.56 dTHG-9 8549 -59.00 3.25 1.00:1.06:0.97:3.55 d THG-10 6725 -56.90 2.37 1.00 :1.01 :0.95 :3.43 SJG 68740 -64.20 54.6 1.00: 1.05 :1.02 : 4.12 dSJG-1 14930 -63.50 16.9 1.00 :0.98 :0.95 :4.09 dSJG-2 9300 -60.30 8.5 1.00 :0.95 : 0.93 : 3.93 HLG 51500 -48.9* 36.5 1.00 : 0.97 : 0.89 : 2.01 dHLG-1 13320 -47.30 5.70 1.00 : 0.97 : 0.85 : 1.98 dHLG-2 9790 -47.6* 4.24 1.00 : 0.95 :0.84: 1.96 19 3305121_i (GHMOM~r) P90582.AU It can be seen from the results of table 2, the molar ratios of GalNAc: GlcUA: Fuc in dTHG, dSJG, dHLG and in their original polysaccharides THG, SJG, HLG were approximately equal to 1:1:1, while the contents of sulfated groups were different, in which the content in SJG/dSJG is higher, and the content in HLG/dHLG is lower. In addition, the optical rotation and intrinsic viscosity between FGAG from the three sources had relatively large differences. For THG/dTHG, the change of the ratio of monosaccharide components in the product produced by the same batches of sea cucumber was small, while in THG and/or dTHG produced by different batches of raw materials, the ratio of the above monosaccharide components changed in a wide range. Generally, the molar ratio of GalNAc: GlcUA: Fuc: -OSO3 was within the range of 1: (1 ± 0.3): (1 ± 0.3): (3.5 + 0.5). It also can be seen from above results that in the product obtained by peroxide depolymerization catalyzed by transitional metal ions, the ratio of the content of fucose to sulfate group changed little, and it is known by 'H-'H COSY analysis that the reducing end of the product was mainly GalNAc, which was substantially identical with the conclusion of the above patent application CN200910110114.0 of the present applicant. Example 2: Spectrum analysis of THG and dTHG Sample: THG, dTHG-6, dTHG-9, with the same sources as Example 1 Control sample: SJG, SJG- 1, with the same sources as Example 1 Detection spectrum: 'H NMR; 'H-'H COSY; 'H-'H TOCSY;H-'H NOESY; 3 C-NMR; DEPT-135 0 ; 'H-"C HSQC; 'H-"C HMBC Detection condition: solvent: D 2 0, 99.9Atom%D (Norell company); internal standard, trimethylsilyl-propionic acid (TSP-d4); temperature, 45 C Instrument: AVANCE AV 400 Superconducting Nucleus Magnetic Resonance Spectrometer (400Mz, Bruker, Switzerland) Spectrum: see Fig.l-Fig.4. Result analysis: (1) Comparison of the spectroscopies of THG and dTHG: Figure 1 shows 1H NMR spectroscopies of THG, dTHG-6, dTHG-9, wherein the water peaks in the spectra of dTHG-6 and dTHG-9 were suppressed. It can be seen from Fig. 1 that THG, dTHG-6 and dTHG-9 had substantially identical signal features, only when the molecular weight was lower, the signal was more distinct. 20 3395121_1 (GI1Mstlcrs) P90562.AU It is seen that the NMR signal features kept stable before and after depolymerization of THG, and the basic chemical structures had no significant change before and after depolymerization of THG. (2) Comparison of the spectroscopies of THG and SJG: With reference to Fig.2-Fig.4, Fig. 2 shows 'H-'H COSY spectroscopies of dTHG (A) and dSJG (B); wherein signals indicated by arrows show the differences in the structure of fucose branches between the two depolymerized FGAG: Fig. A shows the related signals of 3-sulfated fucose of dTHG; Fig. B shows the related signals of 3,4-disulfated fucose of dSJG; signals in panes are respectively the related hydrogen signals of 3-sulfated fucose and 3,4-disulfated fucose; signals in the two circles are the signals of H-4 position of 3,4-disulfated fucose. Fig. 3: shows 3 C NMR and DEPT spectra of THG and SJG. Fig. 4 shows NMR homonuclear/heteronuclear correlation spectra of dTHG-6. It can be seen by summarizing the spectrum data that THG and SJG had substantially the same backbone structure, and the distinct difference between them lied in: as shown in Fig.2, strong signal of 3,4-disulfated fucose of dSJG was weak or did not occur in the spectroscopy of THG; and the signal belonging to 3-sulfated fucose of THG did not occur in the spectroscopy of SJG, indicating that they had significant difference in the substituent type on branches. With reference to Fig.3, carbon signals of GalNAc having and having no sulfate substituent at 6-position occured at about 70 ppm and about 64 ppm respectively. Due to C-6 position of GalNAc being a secondary carbon, its signal peak in DEPT 1350 spectroscopy was a negative peak. It can be seen from Fig.3, there was still small amount of GalNAc (less than 10%) whose hydroxyl groups at C-6 positions was not substituted by sulfate groups in the backbone of THG, while there substantially existed no GalNAc whose C-6 positions was not substituted by sulfate groups in the backbone of SJG. This further demonstrated that there was difference between the substituent groups on branches of THG and SJG. In addition, as can be seen from the existing literature, there was difference in the substituent on branches between THG of present invention and the other known FGAG from sea cucumber. For example, the references (Paulo AS et al., J. Biol. Chem. 1996, 271: 23973; Lubor Borsig et al., J. Biol. Chem. 2007, 282, 14984) and the accompanying 3 C NMR spectroscopy showed that about 35% of GalNAc in FGAG from L. grisea were not substituted by sulfate groups at C-6 position. Reference (Huizeng Fan, et al., Pharmaceutical Journal, 1980, 18(3): 203) showed 21 3395121_1 (GHMnners) P90582.AU that the backbone GalNAc of FGAG from Holothuria leucospilota only had 6-position sulfation and not 4-position sulfation. To sum up, THG of the present invention has chemical structure, which differed from the known structures of FGAG from other species. Firstly, it has different substituent types on fucose branches, namely mainly comprising -2,4-disulfated, -3-sulfated, and -4-sulfated fucose, while had no or little -3,4-disulfated fucose; and its backbone was different to some extend from FGAG from other source. (3) Data and assignment of NMR spectroscopies of dTHG-6: see Table 3 for assignment of the spectroscopy, see Fig.4 for several relevant spectroscopy. Table 3 Data and assignment of 'H/ 3 C-NMR spectroscopies of THG-6 'H Correlated hydrogen shown in hC Correlated hydrogen Position NMR spectroscopies NMR DEPT specto ies assignment [ppm] COSY TOCSY NOESY [ppm] 90/1350 HMQC HMBC 1 4.45 2 2, 3 3 4 ' 102.5 CH I U-4 2 4.04 11, 3 11,31,4 -- 54.1 CH 2# 11, 3 3 3.93 24, 4 21, 4 11, 4, 5 78.2 CH 3 , 2 4 4.82 31, 5 3,5 # , 79.4 CH 4 3 4 6 GaINAc 6 , 6 4 4S6S 5 74"% 4%,6% 4% 5" 6' 6 5 3.9 6' 61 # 6" h 74.6 CR (A) 6 4.23 51, 6'9 5, 6, 6 6# 3r4-, # 69.3 CH 2 6, 5, 6' 6' 4.11 51, 6 51,6 3 6 7 / / / / 177.7 C / 2_ , 8_ 1 3"$ 41 8 2.01 / / '5 25.4 CH 3 8 8 1 4.51 2" 2, 3" [3', 4], 5" 102.5 CH I11 2 3.91 11, 3" 1,' / 55.3 CH 21, 3 3 3.80 24, 4" [2',44] [---] 78.2 CH 3 4 4.76 3, 5 [31,51] [---] 80.4 CH 4 Ga4NA c6 4[, 61 4S 5 3.73 6' 6"' 74.6 CH 5 4 , 6 (A ') 6 3.62 5", 6'" [51 6'1"] [3 , 6,4 .
#6 # 64.1 CR 2 611 6'1 # # ~[3,4,5, 6' 3.69 54, 6 [51 61] 3 , 7 / / / / 177.7 C / 2#, 8# 8 1.98 / /I 4 25.4 CH 3 8 8# 22 3395121_1 (GHMatters) P90562.AU 1 4.42 2 # ' 3 , 5" 106.6 CH I ' ____4 ____ 4
,
5 A-3, C 2 3.57 " 3 ' 4 74.7 CH 2 3 GlcUA 3 3.71 2 , 4 4 1 5 79.9 CH 3 2 4 4 5f (U) 4 3.90 3, 5 #' 2 79.1 CH 3 4 54 5 3.66 4 ' 3, 79.6 CH 5 4 6 / / / / 177.7 C / 4 5 1 5.64 2, 3" 24, U3" 99.2 CH I' U-3# Fuc-2S4 2 4.45 7 _3 1, 3", 4' 1- 76.5 CH 27 7, 3, 44 S 3 4.09 2,4 1, 2, 4 4, 5, 6 69.3 CH 3 1, H~ -*r- ff af ff 4ff 4 4.84 3 5 3 , 5 3 , 5 , 6 83.8 CH 4 5 , 6 (1) 5 4.90 4 6 4 64 3, 44, 6 69.0 CH 5 4 6 1.31 5" 4 ,5" 3, 4, 5 18.6 CH 3 6" 5, 6 1 5.36 24 2 , 3" 2 , U3 4 101.3 CH 14 2 2 4.13 1, 3' 1', 3', 4' l' 73.3 CH 2 # Fue-3S 3 4.60 24 4 14 20, 4 4, 5, 67 83.8 CH 34 4 (11) 4 4.18 3 , 5 3 , 5 3 , 5 , 6 71.9 CH 4' 3# 5 4.47 4,6 4, 6 3,4, 6 69.7 CH 5 3,4 6 1.31 54 4!,5 3 4 4, 5 18.6 CH 3 6 6 1 5.28 2# 2', 3' 2', U3# 101.2 CH I 2 3.80 , 3 1 3 1 71.1 CH 2 1 2 Fuc-4S 3 3.99 2 , 4 1 24, 4 4 , 5 , 6 71.2 CH 3 4 , 5 (111) 4 4.78 3 , 5 3 3, 57, 6T 83.8 CH 4I 54, 6# 5 4.80 4 , 6 44, 6 3 , 4, 6 69.0 CH 5 4 6 1.31 54 5N 3#, 4', 5 18.0 CH 3 6 5#, 6" In the above table, GaINAc4S6S represents 4,6-disulfated-N-acetyl galactosaminyl; GaINAc4S represents 4-sulfated-N- acetyl galactosaminyl; Fuc-2S4S represents 2,4-disulfated fucose; Fuc-3 represents 3-sulfated fucose; Fuc-4S represents 4-sulfated fucose; Fuc-OS represents unsulfated fucose; bracket [] represents data undetermined due to overlapping signals. Example 3: Detection of biological activities of THG/dTHG 3.1 Detection of platelet inducing activity Sample: dTHG-1-dTHG-10, with the same source as example 1 Control sample: dSJG-1~ dSJG-4, with the same source as example 1 Method: New Zealand white rabbit was taken blood from abdominal aorta. The blood was antifreezed with 3.8% sodium citrate (1:9) and centrifugated at 200xg for 8 min to get platelet-rich plasma (PRP), and centrifugated at 1500x g for 10 min to get platelet-poor plasma. Platelet count of PRP is about 4.0 x 10 5 /mm 3 . Bron method (Born GVR. Nature, 1962, 194:927): The effect of the samples on platelet aggregation was detected with platelet aggregometer. In the experiment, physiological saline was used as blank control, and the final concentration of the 23 3395121_1 (GHMatters) P90562.AU sample was 200pg/ml. The experiment was repeated for 3 times and the mean values of maximum degree of platelet aggregation were calculated. Results: see Table 4. Table 4 Comparison of platelet aggregation-inducing activityof different FGAG Sample NS THG dTHG-1 dTHG-2 dTHG-3 dTHG-4 aggreat n rate 2.7 ±4.1 28.1 ±7.8 16.3 ±5.6 12.8 ±4.9 9.3 ±4.8 3.9 ±2.8 Sample dTHG-5 dTHG-6 dTHG-7 dTHG-8 dTHG-9 dTHG-10 ag ratelet 1.9 ± 2.9 1.1 ± 1.3 2.9 ±3.3 2.4 ±3.3 1.2 ±2.5 3.4 ±4.3 Sample SJG dSJG-l dSJG-2 SJG dHLG-1 dHLG-2 Platelet 58.4 ± 13.6 22.7 ± 8.5 5.3 ± 5.5 53.6 ± 17.4 18.3 ± 8.6 4.6 ± 5.4 aggregation rate By comparing the platelet inducing activity of the above FGAG from Thelenota ananas, Stichopusjaponicas and Holothuria leucospilota and the depolymerized product with different depolymerization degrees, it can be found that for original FGAG, the platelet aggregation-inducing activity of THG was far lower than that of SGJ and HLG, indicating that the structural difference of FGAG from different sources had a great effect on the platelet activity. Since the structure difference of THG, SJG and HLG mainly lied in the type difference of sulfated fucose branches, thus it is presumed that the difference of platelet activity attributes to the difference of fucose branches. The type/feature of fucose branches of THG effectively weakened its platelet aggregation activity. From the molecular weight, when the weight average molecular weight of THG was decreased to about 20000Da, the platelet aggregation activity of dTHG disappeared, while only when the molecular weight of SJG and HLG decreased to about 9000-12000Da, the platelet activation effect under high concentration may be avoided. This substantially coincided with the conclusion of the reference (Huizeng Fan, et al., Journal ofBiological Chemistry, 1993, 9(2): 146), indicating that dTHG has better application value than original THG. 3.2 Detection of anticoagulant activity in vitro Sample: dTHG- 1 -ldTHG- 10, with the same sources as Example 1 Control sample: dSJG-l -dSJG-4;with the same sources as Example 1 Low molecular weight heparin sodium (LMWH): 3500-5500Da, 0.4 mlx 4000AxaIU, Anofi Aventis (France) 24 3395121.1 (GHMatters) P90562 AU Reagent: thrombase (Ila): 123 NIH U/mg, Sigma (USA) Chromogenic substrate of thrombase testing(S): 25 mg/vial, HYPHEN BioMed (France) Heparin Cofactor II (HC-II): 100 pg/vial, HYPHEN BioMed (France) Factor VIII (f.VIII): 2001U/vial, Green Cross (China) Biological Products Co., LTD F.VIII Assay Kit: reagents comprise: Reagents: RI: Human Factor X; R2: Activeation Reagent, human Factor IXa, containing human thrombin, calcium and synthetic phospholipids; R3: SXa- 11, Chomogenic substrate, specific for Factor Xa; R4:Tris-BSA Buffer; HYPHEN BioMed (France). Rabbit platelet-poor plasma: Guangzhou Rui Special Biological Technology Co., Ltd. APTT assay kit (gallogen): Shanghai Sun Biological Technology Co., Ltd. Apparatus: microplate reader, Bio-Rad 680 (USA), BICO Two Channel Coagulometer, Minivolt (Italy) Method: (1) Detection of the activity of inhibiting intrinsic factor Xase (f.Xase, Tenase): The detection method established by f.VIII assay kit and f.VIII reagent was used. 30[d of a series of known concentration of THG, dTHG, SJG, LMWH solution or blank control solution (Tris-BSA buffer solution R4) was mixed with 1.0 IU/ml factor VIII (30pl), then sequentially added with reagent kit R 2 (30pl), R, (30pil), and incubated at 37'C for 1min, then added with R 3 (30pl) and incubated at 37 0 C for 1 min, and added with 20% acetic acid (60pl) to stop the reaction and OD 4 o 5 nm was detected. AOD was calculated based on the blank control (R4), and IC 50 of the inhibition of f.Xase of each sample was calculated by the formula provided in the reference (Sheehan J. P. & Walke E. K., Blood, 2006, 107:3876-3882). (2) Detection of HC-II-dependent anticoagulant activity: 50 kl of a series of the concentration of THG, dTHG, SJG, LMWH solution or blank control solution (Tris-BSA buffer solution R4)was added to a 96 well enzyme marking plate, added and mixed with 50pl of 1gmol/L HC-II, incubated at 37'C for 2min, then added with 50pl of 5U/ml Ila and incubated at 37'C for 2min, then added and mixed with 50pl of 2mmol/l CS-0138, incubated at 37*C for Imin, and added with 50% acetic acid (100 l) to stop the reaction and OD 4 0 5 nm was detected. AOD was calculated based on the blank control (R4), and IC 50 of the inhibition of Ila of each sample was 25 3395121_1 (GHMatters) P90562.AU calculated by the formula provided in the reference (Sheehan J. P. & Walke E. K., Blood, 2006, 107.3876-3882). (3) Detection of the activity of extending APTT time: 10 d of a series of the concentration of THG, dTHG, SJG, LMWH solution or blank control solution (Tris-buffer solution R4) was mixed to 180ptl of rabbit plasma, then the APTT time of each sample was detected according to the method of the kit. The drug concentration for multiplying APTT time (double APTT time) was calculated according to the detection result of each sample. Results: see Table 5 and Fig. 5. Table 5 Anticoagulant activitiy of THG/dTHG and control sample IC50(pLg/ml) Potency ratio Drug Potency Molecular go rationrg ratio of Sample weight of concentration anti-f.Xase Mw (Da) Anti-f.Xase Anti-Ila anti-f.Xase-ant for multiplying extension of i-Ia I] A PTT (gg/m I) APTT 21 THG 65820 0.275 1.25 4.54 1.77 8.66 dTHG-l 46070 0.270 1.56 5.78 2.05 7.59 dTHG-2 33260 0.268 1.64 6.11 2.50 9.32 dTHG-3 25380 0.266 1.75 6.58 3.28 12.3 dTHG-4 19650 0.253 1.90 7.51 3.59 14.2 dTHG-5 17150 0.245 2.27 9.26 4.08 16.6 dTHG-6 13950 0.216 2.84 13.1 4.56 21.1 dTHG-7 11580 0.282 3.55 12.6 6.29 22.3 dTHG-8 10260 0.463 4.02 8.68 8.82 19.1 dTHG-9 8549 0.714 5.96 8.34 10.6 14.8 dTHG-10 6725 0.864 6.23 2.17 21.9 7.6 SJG-2 9300 0.683 5.35 7.83 9.7 14.2 LMWH 3500-5500 9.22 6.08 0.66 4.48 0.48 Potency ratio of anti-f.Xase-anti-Ila: anti-Ila IC 5 o(pg/ml) / anti-f.Xase IC 50 (jig/ml) [21 Potency ratio of anti-f.Xase- APTT extension: drug concentration of multiplying APTT (ptg/ml) lanti-f.Xase IC50 (4g/ml) The literatures (GZ. Feuerstein, et al., Arterioscler Thromb Vasc Biol., 1999, 19:2554; CJ Refino, et al., Arterioscler Thromb Vasc Biol., 2002, 22:517) showed that f.IXa inhibitor may substantially inhibit the formation of thrombus under the dosage of without effecting blood APTT and bleeding time, and bleeding tendency of the anticoagulant was associated with the activity of antithrombin. In addition, clinical trial with low molecular heparin had confirmed that with the improvement of potency ratio of anti Xa/anti Ila, its bleeding tendency was significantly decreased (G. Andriuoli et al. Heamostasis, 1985, 15: 324). Based on above 26 3395121_1 (GHMatters) P90562.AU knowledge and the relationship between molecular weight of dTHG and its potency on different blood coagulation factor targets, appropriate depolymerization of THG may eliminate platelet aggregation-inducing activity of FGAG, but also produce potency ratio of anti f.Xase and HC-II-dependent anti Ila activity as high as possible, and/or potency ratio of anti f.Xase and /or extending APTT activity as high as possible. The results of Table 5 showed that THG and/or dTHG had the activities of extending APTT time, inhibiting intrinsic f.Xase and HC-II dependent antithrombase. Depolymerization of THG may eliminate its platelet inducing activity, but the depolymerization degree had different effect on its activities of anti f.Xase, anti Ila (HC-II-dependent) and the extension of APTT. Firstly, when the molecular weight (Mw) of dTHG was decreased to 20000Da, its platelet inducing activity disappeared completely, and when Mw was decreased to 6000Da, it still had anticoagulant activity (extending APTT time). Obviously, dTHG in the molecular weight range possibly had better application value compared to original THG. Secondly, both THG and dTHG had strong activity of inhibiting endogenous f.Xase. Under the condition mentioned in Example 3.2 of the present invention, dTHG (or THG) with a weight-average molecular weight of about 8000-70000Da generally had an IC 50 of inhibiting f.Xase of less than about 0.1p mol/L (less than about I tg/ml), and exhibited an enhanced tendency with the increase of the molecular weight. However, the tendency had large difference in different molecular weight range. When the molecular weight was less than about 12000Da, the f.Xase-inhibitory activity of dTHG may distinctly weakened with the decrease of the molecular weight; while when the molecular weight was not less than about 12000Da, the IC 50 is in the range of about 0.2ptg/ml-0.3pg/ml, based on molar concentration, the IC 50 may slightly decrease with the increase of the molecular weight; but based on mass concentration, the IC 50 substantially did not change with the molecular weight, or more exactly, it slightly increased with the increase of the molecular weight. As a whole, when dTHG (or THG) had a molecular weight of no less than about 10000 Da, its endogenous f.Xase-inhibitory activity may maintain constant to some extent. The results of Table 5 also showed that THG/dTHG has activities of HC-II dependent antithrombin and extending APTT time. For dTHG (or THG) with a molecular weight of about 8000-70000 Da, both the activity of HC-II-dependent 27 3395121_1 (GHMatleIs) P90582 AU antithrombin and the activity of extending APTT time enhanced with the increase of the molecular weight, which were manifested in the liner decrease of IC 5 0 of HC-II-dependent antithrombin activity and drug concentration of multiplying APTT time with the increase of logarithm value of molecular weight. It is concluded by summarizing the above results according to the present invention that when dTHG had a molecular weight of more than about 1 000ODa, especially about 12000Da, the molecular weight had an effect on extending APTT time and HC-II-dependent antithrombin activity of dTHG, which was much larger than the effect on inhibiting endogenous f.Xase activity. For the convenience of description, "potency ratio of anti-f.Xase-anti-IIa" and "potency ratio of anti-f.Xase- APTT extension" were defined in Table 5 to reflect the features of anticoagulant activity of THG/dTHG with different molecular weight. As a whole, for dTHG with a molecular weight of no less than 1000Da, the lower the molecular weight was, the higher the potency ratio of the f.Xase inhibitory activity to HC-II-dependent antithrombin activity or APTT time extension was. When the molecular weight was more lower (less than about I 000ODa), the potency ratio exhibited a decreased tendency. The results of Table 5 also showed that LMWH had a relatively weak anti-f.Xase activity, and platelet induction activity of dSJG limited the application of the product with a stronger acticity and/or higher "potency ratio". Based on the features of (1) eliminating platelet induction activity, (2) achieving a high anti-Xase/anti-IIa potency ratio as possible, and/or (3) achieving a high anti-Xase/APTT extension potency ratio, the present invention comprehensively pondered the relationship between the molecular weight of dTHG and f.Xase inhibitory activity, HC-II-dependent antithrombin activity, APTT time-extending activity and platelet effect. The dTHG selected according to the present invention may have a weight-average molecular weight (Mw) of about 8000-20000Da, preferably about 10000- 1 8000Da, more preferably about 12000-16000Da. 3.3 Resistance of dTHG to venous thrombosis Sample: dTHG-6, with the same sources as Example 1; Low molecular weight heparin sodium (LMWH): 3500-5500Da, 0.4 mix 4000AxaIU, Anofi-Aventis Method: (1) Venous thrombosis in rabbit: A male New Zealand white rabbit was narcotized with pentobarbital, the bilateral jugular veins were separated and respectively placed two ligation sutures at 2cm segment, and intravenously injected 28 3395121_1 (GHMntters) P90582.AU with recombinant human tissue factor at a dose of lng/kg. The proximal end and distal end of the vein were ligated after 5min, and the blood vessel was longitudinally cut after 15min to take out thrombus. Residual blood was sucked with a filter paper, and wet weight of the thrombus was weighed. Test drugs (dTHG-6, LMWH) and control solvent (physiological saline, NS) were all administered subcutaneously before 2 hours of vein ligation. (2) Detection of bleeding time: A SD rat was narcotized with pentobarbital (30 mg/kg ip), and intravenously injected with dTHG-6 or LMWH. After 15 seconds, its tail was cut off (5mm from the end of the tail). Bleeding from the amputated tail was sucked at every 15 second with filter paper. No bleeding for continuous 1 min was considered as bleeding-stopping. Results: (1) Venous thrombosis in rabbit: In the model of venous thrombosis under the conditions of ligation and hyperviscosity, subcutaneous injection of dTHG-6 at 4.5, 9, 18mg/kg could substantially inhibit thrombosis, and the inhibition rate ranged about 35% to about 70%. The inhibition activity exhibited distinct dose-effect relationship (Table 6). In this experiment, had the inhibition rate of LMWH at 7201U/kg on venous thrombosis was about 56% Table 6 Effect of THG on jugular vein thrombosis GMup n Weight of thrombus (mg) Inhibition rate (%) Model group 10 152.8 ±13.6 - THG 4.5 mg/kg 10 98.3 ± 25.6 * 35.7 9 mg/kg 10 56.4 ± 20.6 * 63.1 18 mg/kg 10 45.7 ± 26.8 * 70.1 LMWH 720 IU/kg 10 67.3 ± 21.2 * 56.0 Compared with the model group: *P<0.05, **P<0.01 (2) Detection of bleeding time: The detection results of the effect of subcutaneous injection of dTHG-6 or LMWH on bleeding time in rat were shown in Fig.6. The experimental results showed that compared with LMWH, under the similar anticoagulant and antithrombotic disage, dTHG-6 had a lower effect on bleeding time. 29 3395121_1 (GHMatters) P90582.AU
Claims (10)
1. A depolymerized glycosaminoglycan from Thelenota ananas and a pharmaceutically acceptable salt thereof, characterized in that said depolymerized glycosaminoglycan from Thelenota ananas is a depolymerized product of fucosylated glycosaminoglycan from Thelenota ananas, which has a weight average molecular weight of 8000-20000Da, has monosaccharide components of N-acetyl galactosamine, glucuronic acid, fucose and their sulfates, and has a molar ratio of N-acetyl galactosamine : glucuronic acid : fucose : the sulfate groups expressed as -OS0 3 of 1: (1±0.3): (1+0.3): (3.5±0.5).
2. The depolymerized glycosaminoglycan from Thelenota ananas and a pharmaceutically acceptable salt thereof according to claim 1, characterized in that in said monosaccharide components, the sulfated fucose comprises 3-sulfate fucose.
3. The depolymerized glycosaminoglycan from Thelenota ananas and a pharmaceutically acceptable salt thereof according to claim 1, characterized in that said depolymerized glycosaminoglycan from Thelenota ananas has a weight average molecular weight of 10000~18000Da.
4. A method for preparing a depolymerized glycosaminoglycan from Thelenota ananas and a pharmaceutically acceptable salt thereof, according to claim 1, comprising the steps of: 1) extracting and obtaining fucosylated glycosaminoglycan from the body wall of Thelenota ananas; 2) depolymerizing the fucosylated glycosaminoglycan obtained in step 1) by peroxide depolymerization to obtain an depolymerized fucosylated glycosaminoglycan; 3) collecting and purifying the depolymerized fucosylated glycosaminoglycan with a desired molecular weight.
5. The method according to claim 4, characterized in that said fucosylated glycosaminoglycan from Thelenota ananas is depolymerized in an aqueous medium in the presence of a catalyst to obtain the depolymerized fucosylated glycosaminoglycan, said catalyst is a catalyst containing a metal ion selected from the group consisting of the fourth period transition metals in the periodic table.
6. The method according to claim 5, wherein said catalyst containing a metal ion selected from the group consisting of the fourth period transition metals in the 3395121_1 (GHMattes) P90582.AU periodic table is an inorganic salt or an organic salt of Cu', Cu2+, Fe 2 +, Fe3+, Cr 3 *, Cr 2 0 7 , Mn 2+, Zn2+, Ni 2 +, or the combination thereof.
7. A pharmaceutical composition comprising the depolymerized glycosaminoglycan from Thelenota ananas or a pharmaceutically acceptable salt thereof according to any one of claims I to 3, and pharmaceutically acceptable excipients.
8. The pharmaceutical composition according to claim 7, characterized in that said pharmaceutically acceptable salt is sodium, potassium or calcium salt.
9. The pharmaceutical composition according to claim 7 or 8, characterized in that said pharmaceutical composition is formulated into freeze-dried powder for injection.
10. A use of the pharmaceutical composition according to claim 7 or 8 in preparing medicines for the prevention and/or treatment of thrombotic diseases. 2
3395121.1 (GHMatters) P90582.AU
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN200910109861A CN101724086B (en) | 2009-11-25 | 2009-11-25 | Oligomerization pineapple ginseng glycosaminoglycan and preparation method thereof |
CN200910109861.2 | 2009-11-25 | ||
PCT/CN2010/001678 WO2011063595A1 (en) | 2009-11-25 | 2010-10-25 | Depolymerized glycosaminoglycan from thelenota ananas and preparation method thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
AU2010324437A1 true AU2010324437A1 (en) | 2012-07-19 |
AU2010324437B2 AU2010324437B2 (en) | 2014-12-18 |
Family
ID=42445696
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2010324437A Ceased AU2010324437B2 (en) | 2009-11-25 | 2010-10-25 | Depolymerized glycosaminoglycan from Thelenota ananas and preparation method thereof |
Country Status (4)
Country | Link |
---|---|
US (1) | US8809300B2 (en) |
CN (1) | CN101724086B (en) |
AU (1) | AU2010324437B2 (en) |
WO (1) | WO2011063595A1 (en) |
Families Citing this family (27)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101724086B (en) * | 2009-11-25 | 2012-09-26 | 深圳海王药业有限公司 | Oligomerization pineapple ginseng glycosaminoglycan and preparation method thereof |
CN102247401A (en) * | 2011-05-05 | 2011-11-23 | 中国科学院昆明植物研究所 | Low molecular weight glycosylated chondroitin sulfate and its purpose in preparation of anti-HIV-1 medicament |
CN102329397B (en) * | 2011-10-19 | 2014-04-09 | 中国科学院昆明植物研究所 | Fucosylated glycosaminoglycan derivative and preparation method thereof |
CN102558389B (en) * | 2011-12-22 | 2013-10-02 | 中国科学院昆明植物研究所 | Low molecular weight carboxyl-reduced derivatives of fucosylated glycosaminoglycans and preparation method and applications of low molecular weight carboxyl-reduced derivatives |
CN103285031B (en) * | 2012-03-05 | 2015-09-09 | 上海开润生物医药有限公司 | The application of depolymerization glycosaminoglycan extracted from sea cucumber in preparation control thromboembolic disorders medicine |
CN103417565B (en) * | 2012-05-17 | 2016-05-25 | 上海开润生物医药有限公司 | The application of depolymerization glycosaminoglycan extracted from sea cucumber in preparation control thrombotic diseases medicine |
CN103830263A (en) * | 2012-11-26 | 2014-06-04 | 上海开润生物医药有限公司 | Application of depolymerized holothurian glycosaminolycan in medicine preparation |
CN103830264A (en) * | 2012-11-26 | 2014-06-04 | 上海开润生物医药有限公司 | Application of depolymerized holothuria glycosaminoglcan in preparing medicine for preventing and controlling thrombotic diseases |
CN103869002B (en) * | 2012-12-11 | 2015-05-27 | 深圳海王药业有限公司 | Analysis method for determining oligomerization thelenota ananas glycosaminoglycan content |
CN102993323A (en) * | 2012-12-30 | 2013-03-27 | 青岛市市立医院 | Method for extracting and purifying glycosaminoglycans (GAGs) from stichopus japonicus |
CN103044564A (en) * | 2012-12-30 | 2013-04-17 | 青岛市市立医院 | Preparation method for holothuria leucospilota glycosaminoglycans |
CN103044565A (en) * | 2012-12-30 | 2013-04-17 | 青岛市市立医院 | Method for extracting black sea cucumber glycosaminoglycan |
CN102993327A (en) * | 2012-12-30 | 2013-03-27 | 青岛市市立医院 | Method for preparing black holothurian glycosaminoglycan |
CN102993326A (en) * | 2012-12-30 | 2013-03-27 | 青岛市市立医院 | Method for extracting and purifying glycosaminoglycans (GAGs) from thelenota ananas |
CN103030706A (en) * | 2012-12-30 | 2013-04-10 | 青岛市市立医院 | Method for extracting glycosaminoglycan from thelenota ananas |
CN103087207A (en) * | 2012-12-30 | 2013-05-08 | 青岛市市立医院 | Method for extracting holothuria leucospilota glycosaminoglycans |
EP2980103B1 (en) | 2013-03-26 | 2020-11-04 | Jiuzhitang Co., Ltd. | Low molecular weight glycosaminoglycan derivative, pharmaceutical composition thereof, preparation method therefor and use thereof |
CN103214591B (en) | 2013-04-12 | 2015-11-04 | 中国科学院昆明植物研究所 | A kind of lower molecular weight osamine polysaccharid derivative of the talose or derivatives thereof that dewaters containing end 2,5- |
CN104147040A (en) * | 2013-05-13 | 2014-11-19 | 上海开润生物医药有限公司 | Application of holothuria glycosaminoglcan in preparation of medicines for preventing and treating thromboembolism disease |
CN103788222B (en) | 2014-01-08 | 2016-08-31 | 中国科学院昆明植物研究所 | Substituted oligomeric glycosaminoglycans of Fuc3S4S and preparation method thereof |
TWI769176B (en) * | 2016-09-07 | 2022-07-01 | 瑞瑟勒綜合技術協會 | Biosynthetic heparin |
JP6980018B2 (en) | 2017-01-10 | 2021-12-15 | 九芝堂股▲ふん▼有限公司Jiuzhitang Co., Ltd. | Oligosaccharides that inhibit the endogenous tennase complex, their production methods and uses |
CN108344875B (en) * | 2017-01-22 | 2021-11-02 | 上海长岛生物技术有限公司 | Method for improving sensitivity of reagent for activating partial thromboplastin time to heparin and application |
AU2020209822A1 (en) | 2019-01-15 | 2021-09-09 | Optimvia, Llc | Engineered aryl sulfate-dependent enzymes |
EP3997238A1 (en) | 2019-07-09 | 2022-05-18 | Optimvia, LLC | Methods for synthesizing anticoagulant polysaccharides |
CN114252546A (en) * | 2020-09-23 | 2022-03-29 | 牡丹江友搏药业有限责任公司 | Method for determining content of low-molecular-weight fucosylated glycosaminoglycan |
CN114252436A (en) * | 2020-09-23 | 2022-03-29 | 牡丹江友搏药业有限责任公司 | Method for identifying natural fucosylated glycosaminoglycan |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP3455783B2 (en) * | 1995-12-20 | 2003-10-14 | 大鵬薬品工業株式会社 | Intimal thickening inhibitor |
EP1551852A4 (en) | 2002-04-25 | 2007-03-21 | Momenta Pharmaceuticals Inc | Methods and products for mucosal delivery |
AR043110A1 (en) * | 2004-02-04 | 2005-07-20 | Syntex Sa | LOW MOLECULAR HEPARINE SALT WITH USEFUL TRIETHANOLAMINE AS A THERAPEUTIC-ANTITROMBOTIC AGENT OF TOPICAL ADMINISTRATION, PROCEDURES TO PREPARE SALES, PROCESS FOR THE ELIMINATION OF HYPROSCOPICITY SALT IN PHAROSERIC ACIACIACEUTIC USE IN TOP THERAPY PAPER USE |
JP2007008899A (en) * | 2005-07-04 | 2007-01-18 | Mie Univ | Vascularization inhibitor |
CN100525777C (en) * | 2007-05-14 | 2009-08-12 | 张登科 | Depolymerization glycosaminoglycan extracted from sea cucumber composition and its preparation method and application |
CN101451157B (en) * | 2008-12-25 | 2012-09-05 | 大连海晏堂生物有限公司 | Method for preparing low molecular weight sea cucumber polysaccharide |
CN101724086B (en) | 2009-11-25 | 2012-09-26 | 深圳海王药业有限公司 | Oligomerization pineapple ginseng glycosaminoglycan and preparation method thereof |
-
2009
- 2009-11-25 CN CN200910109861A patent/CN101724086B/en active Active
-
2010
- 2010-10-25 WO PCT/CN2010/001678 patent/WO2011063595A1/en active Application Filing
- 2010-10-25 AU AU2010324437A patent/AU2010324437B2/en not_active Ceased
- 2010-10-25 US US13/512,142 patent/US8809300B2/en active Active
Also Published As
Publication number | Publication date |
---|---|
WO2011063595A1 (en) | 2011-06-03 |
CN101724086B (en) | 2012-09-26 |
CN101724086A (en) | 2010-06-09 |
AU2010324437B2 (en) | 2014-12-18 |
US8809300B2 (en) | 2014-08-19 |
US20120270834A1 (en) | 2012-10-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2010324437B2 (en) | Depolymerized glycosaminoglycan from Thelenota ananas and preparation method thereof | |
US4500519A (en) | Mucopolysaccharides having biological properties, preparation and method of use | |
US10689463B2 (en) | Fuc3S4S substituted oligoglycosaminoglycan and preparation method thereof | |
CN102329397B (en) | Fucosylated glycosaminoglycan derivative and preparation method thereof | |
US4804652A (en) | Mucopolysaccharides having biological properties, preparation and application thereof as drugs | |
EP2794666B1 (en) | Use of chemically modified heparin derivates in sickle cell disease | |
EP0337327A1 (en) | Process for the preparation of new oligosaccharide fractions by controlled chemical depolimerization of heparin | |
EP0048231A1 (en) | Oligosaccharides having selective anticoagulation activity | |
WO2000006608A1 (en) | Novel glycosaminoglycan and drug compositions containing the same | |
US10494452B2 (en) | Low-molecular-weight glycosaminoglycan derivative containing terminal 2, 5-anhydrated talose or derivative thereof | |
Spadarella et al. | From unfractionated heparin to pentasaccharide: Paradigm of rigorous science growing in the understanding of the in vivo thrombin generation | |
JP2628507B2 (en) | EDTA-free heparin, heparin fractions and fragments, processes for their preparation and pharmaceutical compositions containing them | |
CN108285498B (en) | Oligosaccharide compound for inhibiting endogenous coagulation factor X enzyme complex and preparation method and application thereof | |
KR970000528B1 (en) | New sulfated polysaccharide, pharmaceutically acceptable salts thereof, preparation thereof and drug containing the same as active ingredient | |
CN109251255B (en) | Fucosylated chondroitin sulfate FCShmAnd preparation method and application thereof | |
US11202797B2 (en) | Mixture of fucosylated chondroitin sulfate oligosaccharides and method for rapidly producing the same | |
Viskov et al. | Isolation and characterization of contaminants in recalled unfractionated heparin and low-molecular-weight heparin | |
EP3569608A1 (en) | Oligosaccharide compound for inhibiting endogenous coagulation factor x-enzyme complex, and preparation method therefor and uses thereof | |
CN115448994A (en) | Neutralization anticoagulant low molecular weight heparin and preparation method and application thereof | |
Ventre | FROM UNFRACTIONATED HEPARIN TO PENTASACCHARIDE: PARADIGM OF RIGOROUS SCIENCE GROWING IN THE UNDERSTANDING OF THE IN VIVO THROMBIN GENERATION | |
EP0351809A2 (en) | Blood anticoagulant |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
FGA | Letters patent sealed or granted (standard patent) | ||
MK14 | Patent ceased section 143(a) (annual fees not paid) or expired |